Dual Modulation Two-dimensional Gas Chromatograph Time Of flight (2D GC-

TOF-MS) with Three dimensional Plot – A new Approach

Summery:
The GCXGC, which is known as Two Dimensional GC (2D GC) is the most powerful
Gas Chromatography system for the analysis of complex samples like essential oils,
petroleum products, residual pesticides in food, etc., which can be coupled with MS and
provide the all information in single analysis. The complexity in qualitative analysis
because of co-eluting components are not at all a problem now.

Introduction:
Chromatography is a separation technology to separate individual components from a
mixture. We all know, when the separation relies on a different bulk parameter, such as
the distribution ratio between the stationary phase and the gas phase in GC, the
compound’s partition coefficient, k, is defined by the familiar relationship k=Cs / Cm,
where as Cs is the concentration of the compound in the stationary phase and Cm is the
concentration of the analyte in the mobile phase. Factors that influence this parameters
include the nature of the stationary phase (chemical composition, size, shape, polarity,
etc.), and instrumental parameters such as temperature, pressure. All of these parameters
contribute towards the partition coefficient, which can be viewed as the bulk parameter
that provides the separation. If the multiple components have the same value of k, they
will not separate using that particular parameter, we have to select different stationary
phase.
When dealing with mixture of components, it is often impossible to separate all of them
using any single separation dimension. Refer the bellow chromatogram of Mint on a non-
polar column.

7
8+9+10
11

16

18
17
14
12
4
2 3
13 15
1 6 19

20 25 30 35 40 45 50 55
time (s)

1) a-Pinene 6) g-Terpinene 11) Menthol 16) Menthyl acetate

2) Sabinene Hydrate 7) Menthone 12) Terpinenene-4-ol 17) b-Cariophyllene
3) b-Pinene 8) i-Menthone 13) a-Terpineol 18) D-Germacrene
4) 1,8-Cineole 9) Menthofurane 14) Pulegone 19) Viridifluorol
5) Limonene 10) neo-menthol 15) Piperitone

Capillary column RTX-5 used to separate the above components. The separation of
component 8, 9 & 10 is not there.

The same sample analyzed on a polar capillary column Mega-Wax.

9 17
7 13
14

15
10 16
6 20
11 18

2
1 8 19
3 5 12 21
22
4

15 20 25 30 35 40 45 50 55 60 65 70 75
time (s)

1) a-Pinene 7) 1,8-Cineole 13) Menthyl acetate 19) a-Terpineol

2) b-Pinene 8) g-Terpinene 14) neo-menthol 20) Germacrene
3) Sabinene Hydrate 9) Menthone 15) Terpinenene-4-ol 21) Piperitone
4) Myrcene 10) Menthofurane 16) b-Cariophyllene 22) Viridifluorol
5) p-Cymene 11) i-Menthone 17) Menthol
6) Limonene 12) Linalool 18) Pulegone

Here the separation between 8,9 & 10 is is very nice, but we loose the resolution between
17 & 18, 20 & 21.

This is a very simple example. Most of the times to solve this problem we do the multiple
analysis by changing the polarity of the column ( so the different k factor). Or we use the
heart-cut analysis, where a particular portion of a chromatogram cuts and introduce to a
different polarity column. This heart-cut analysis is trouble some and lots of trial has to
take. Because of the column bleeding, the retention time changes and so have to change
the time of “Heart-Cut”.

Following is an schematic example of heart-cut analysis.

In the heart-cut analysis, the component injected first into a non-polar column which
separate the components as per boiling points. But the components which are very close
to the boiling points co-clutes together which switched over to the polar column and can
be separated. However, the number of heart-cuts from the first coumn that can be
analyzed by a second column is limited by the analysis and recovery times of the second
column before each subsequent heart-cut cycle. Peaks that are eluted from the first
column while the second column is actively analyzing a previously sampled heart-cut do
not benefit from separation in the second column and are sent to the first detector instead.
One solution for this problem of peaks queuing up and waiting for the second column to
finish a separation is to use more than one secondary column. Or to make the secondary
separation faster so that queuing peaks do no t accumulate too much during each
secondary separation. That’s why, the GCXGC concept is now widely using in world
market.

GCXGC:
Comprehensive Two-Dimensional gas chromatography has been proved to be clearly
advantageous for qualitative analysis because of its enhanced separation power and its
peculiar capability of ordering the compounds according to their chemical and physical
properties with sophisticated software to handle GCxGC data for quantitation.
In the GCXGC technique, the first column is always a conventional capillary column of
25 mtr. Length and of 0.25mm ID and the second column is of very short length (from
2mt. to 5mt.) with 0.1mm ID. In most of the cases the first column is non-polar and the
second column is polar. But there is a possibility to reverse the polarity if the analytical
requirement is there.
The most prevalent methods today involve some form of thermal modulation at the
juncture of columns 1 and 2.

The above figure illustrates the main principles of GCXGC operation. The mechanical
and electronic arrangements blow the liquid CO2 or N2 with a prefixed and adjustable
time interval over a section of beginning of column 2 (known as modulation). When the
cooling flow turned off, the cooled column section rapidly reheats from circulating hot
oven air.
When the cooling is applied, peaks that are eluted from the first column are trapped in
place at the beginning of the second column. So the band of the peak compressed. When
heated, the peaks are released and begin to move again in a narrow band to start the
secondary separation. If the system is with a single point modulation, during the heating
stage, if any new peaks enters in to the second column will not be trapped and will over
lap with the components which getting separated by the second column. This problem
can be eliminated with 2 point modulation.
 stretched column

self-aligned CO2 jets

The above picture is the inside of the GC oven with 2 point modulation.

Thus a series of rapidly repeating injections are performed efficiently onto the secondary
column with slices trapped off the first column.

Software:
The visualization of the data obtained from the GCXGC can not be become apparent until
some data manipulation is performed. The most common data transformation is the
construction of 2D plot in which one axis represents the separation on the first column
and other axis represents the separation on second column. A contour plot using elevation
lines or color coding represents the signal intensity. A third dimension can be added
optionally by making the z axis proportional to the signal intensity.
 linear scale 3D rendering

log scale apex plot

To generate the reconstructed 2-D plot, the software use the timing of the thermal
modulators to locate the start of each secondary column run in the raw data stream. The
raw data stream is sliced into multiple sub chromatograms that represent each of the
secondary runs. Please refer the bellow diagrams.
These slices can be assembled side by side to produce a two or three dimensional map of
the overall separation. The map is very useful because it shows relationships between
groups of peaks in both the primary and secondary column separations.
This technique is very useful for the analysis of petroleum products, essential oils,
residual pesticides, biomedical, forensic studies, environmental analysis, geochemistry,
etc, where to get the separation of each and every component multiple columns are
required.
This GCXGC can be used with FID, FFPD, SCD and also with the MS. Different
Chromatographers in world has demonstrated its capability and useful ness for the
analysis OF LCO, PIONA, Wide range of hydrocarbons (C6 to C40), crude oil, jet fuels,
etc. in petroleum industries, different essential oils, residual pesticides in fruits (insub
ppm level using FID), etc.
The problem with conventional detector like FID, FPD or ECD is that these detectors can
not use for unknown components as ONLY GC is a relative technology and so useonly
for known component quantification. But the power of 2D GC is to analyze for unknown
components and so it should be coupled with TOF (Time Of Flight) Mass Spectrometry.
Using this combination, it is possible to detect each and every volatile organic component
present in the sample by doing library matching of the spectra obtained from the
component analyzed.

The advantages of 2D GC-TOF:

1. Analysis is faster.
2. Easy to separate each and every component present in the sample.
3. No need of multiple hurt cut analysis.
4. Analysis by single injection for the complex samples for which multiple column
and injection required in conventional GC.
5. Easy identification without using any standards.
6. No peak over lapping.
7. Wide dynamic range.
8. Possible to do group identification along with individual peak identification.

Application area:
The following area of application is best for 2D GC.
1. Residual pesticide in soil, food products and water.
2. Petrochemical fields for details hydrocarbon analysis.
3. Plan extract for essential oils, aromas, low molecular weight drugs and pesticides.
4. PAH analysis in soil and water.
5. Environment analysis for VOCs and other compoents which are not good for
health and environment.